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Purification Solutions for Plant-Derived Recombinant Collagen

1. Overview of Plant-Derived Recombinant Collagen

As a high-profile biomanufacturing product in recent years, plant-derived recombinant collagen has witnessed robust industry growth, with continuous entry of new manufacturers and product launches. An increasing number of collagen production workflows have shifted toward plant-based recombinant expression systems.

Compared with traditional animal-derived collagen and other recombinant expression platforms, plant-derived recombinant collagen presents distinct advantages in biosafety, structural integrity, precise post-translational modification, large-scale production cost efficiency and ethical compliance, making it an optimal alternative.

2. Major Challenges and Corresponding Solutions for Purification

Plant-derived recombinant collagen is categorized into two types: truncated recombinant collagen and full-length recombinant collagen. The core purification challenges stem from the complex impurities inherent in plant hosts, including abundant polyphenols, pigments and polysaccharides. These contaminants tend to interfere with target binding and cause collagen degradation, while plant gum residues are also difficult to remove thoroughly. In particular, full-length collagen exists as an elongated trimer macromolecule with a length of 300 nm and a width of 1–2 nm, leading to great difficulties in target capture.

Advantages for Full-Length Plant-Derived Collagen

High binding capacity: The super-macroporous media achieves a dynamic binding capacity of 5 mg/mL for recombinant human collagen, outperforming conventional non-super-macroporous media.

High purity: It specifically captures full-length collagen with intact triple-helix structure. The product shows high purity verified by enzymatic digestion and electrophoresis, complying with purity specifications for subsequent formulation processing.

Advantages for Truncated Plant-Derived Collagen

High turbidity tolerance: Compatible with feedstock with a turbidity up to 500 NTU after homogenization and centrifugation, allowing particulate impurities to flow through directly.

High viscosity tolerance: Direct loading is feasible for feedstock with a viscosity of approximately 1.0 mPa·s affected by plant gum.

3. Downstream Purification Processes for Plant-Derived Collagen

Purification Process for Full-Length Plant-Derived Collagen

Transgenic plant leaves/seeds → Liquid nitrogen grinding → Acidic extraction → Salting out / Ultrafiltration → Composite cation exchange capture / Butyl hydrophobic interaction capture → Enzymatic digestion → Composite cation exchange polishing → Formulation

Application Case (Full-Length Recombinant Human Collagen with Intact Triple-Helix Structure)

Challenge: Conventional non-super-macroporous media feature low binding capacity and poor capture efficiency for target collagen.

Solution: Adopt GPB super-macroporous hydrophobic media for target capture. This polymeric media features tunable pore sizes, and the GP series super-macroporous media has a maximum pore size of 1000 nm.

Results:

1.Superior binding capacity: A dynamic binding capacity of 5 mg/mL for recombinant human collagen is realized, far exceeding that of non-super-macroporous counterparts.

2.Excellent purity: The media selectively captures full-length collagen with native triple-helix structure. High product purity is confirmed by enzymatic digestion and electrophoresis, meeting relevant quality requirements.

Purification Process for Truncated Plant-Derived Collagen

Transgenic plant leaves/seeds → Liquid nitrogen grinding + Acidic extraction → Salting out / Ultrafiltration → Composite cation exchange capture / Butyl hydrophobic interaction capture → Enzymatic digestion → Composite cation exchange polishing → Formulation

Application Case (Truncated Recombinant Human Collagen)

Challenge: Plant hosts contain complex impurities such as polyphenols, pigments and polysaccharides, which interfere with ligand binding and induce collagen degradation. In addition, plant gum cannot be completely removed during purification.

Solution: A three-step purification workflow using HPP-90, MMC FF and CPQHL-60 to reach the target purity of 98%.

Results:

1.HPP-90 tolerates viscous feedstock and removes the majority of polyphenols, pigments and polysaccharides.

2.MMC FF performs fine polishing of truncated collagen.

3.CPQHL-60 eliminates host cell derived (HCD) residual impurities.

4.The overall recovery rate exceeds 70%, and the final product purity is above 98%

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Purification Solutions for Plant-Derived Recombinant Collagen

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